细粒棘球绦虫TSP8基因在其不同发育阶段的差异表达及生物信息学分析  

Differential Expression and Bioinformatics Analysis of TSP8 Gene of Echinococcus granulosus at Different Developmental Stages

作　　者：王炜烨 王云菲 陆宝燕 马勋[1] 张艳艳[2] 孟季蒙[2] 王正荣[2] 薄新文[1,2] WANG WeiYe;WANG YunFei;LU BaoYan;Ma Xun;ZHANG Yanyan;MENG Jimeng;WANG Zhengrong;BO Xinwen(College of Animal Science and Technology,Shihezi 832000,China;State Key Laboratory of sheep Genetic Improvement and Healthy Production/Institute of Animal Husbandry and Veterinary,Xinjiang Academy of Agricultural and Reclamation Sciences,Shihezi 832000,China)

机构地区：[1]石河子大学动物科技学院,石河子832000 [2]新疆农垦科学院畜牧兽医研究所省部共建绵羊遗传改良与健康养殖国家重点实验室,石河子832000

出　　处：《中国动物传染病学报》2022年第1期157-165,共9页Chinese Journal of Animal Infectious Diseases

基　　金：国家重点基础研究发展计划(973计划)(2015CB150300);国家自然科学基金(31860701,31360608);新疆生产建设兵团国际科技合作(2017BC003)。

摘　　要：为研究细粒棘球绦虫(Echinococcus granulosus,E.g)TSP8基因的分子特性以及其在虫体不同发育阶段的差异表达,根据NCBI GenBank数据库中的TSP8基因设计1对特异性引物,对TSP8基因进行PCR扩增,将PCR产物克隆到pMD19-T载体后测序,通过生物信息学软件分析预测TSP8基因的结构与功能,并通过SYBR GreenⅠqRT-PCR方法分析TSP8基因在虫体原头蚴、包囊壁以及成虫mRNA相对转录情况。结果显示:成功克隆到细粒棘球绦虫TSP8基因序列,与细粒棘球绦虫跨膜蛋白(GenBank登录号:CDS17460.1)同源性为100%;进一步生物信息学分析显示,TSP8基因全长669个核苷酸,编码222个氨基酸,理论等电点为5.06,为稳定蛋白分子;TSP8基因编码的氨基酸序列共含有5个跨膜区域,含有4个优势B抗原表位;qRT-PCR结果显示TSP8基因在原头蚴及成虫阶段均有表达,差异显著,具有统计学意义(P<0.05)。本研究对细粒棘球绦虫TSP8基因的分子特性进行了初步研究,为进一步揭示其生物学功能奠定了基础。In order to study the molecular characteristics of the TSP8 gene of Echinococcus granulosus and its differential expression at different developmental stages,a pair of specifi c primers was designed according to the TSP8 gene in the NCBI GenBank database,and the TSP8 gene was amplifi ed by PCR.The PCR product was cloned into the pMD19-T vector and sequenced for bioinformatics analysis.The relative transcription level of TSP8 gene in the hydatid protoscolex,cyst and adult mRNA was analyzed by SYBR GreenⅠqRT-PCR.The results showed that the TSP8 gene sequence of E.granulosus was successfully cloned.The gene sequence showed 100%identity with the TSP8(CDS17460.1)as reported.Further bioinformatics analysis showed that the TSP8 gene was 669 nucleotides in length and encoded 222 amino acids.The theoretical isoelectric point was 5.06,which was a stable protein.The amino acid sequence encoding the TSP8 gene contained fi ve transmembrane regions,presumably contained four dominant B epitopes.Additionally,the qRT-PCR results showed that the TSP8 gene was expressed in both the protozoa and adult stages with signifi cant difference(P<0.05).In conclusion,the present study revealed the molecular characteristics of the TSP8 gene of E.granulosus and provided a foundation for further research on its biological functions.

关 键 词：细粒棘球绦虫 基因克隆 差异表达 生物信息学分析 

分 类 号：S852.734[农业科学—基础兽医学]